# 603: ANCALOMICROBIUM MEDIUM


## Main sol. 603

0.25	g	(NH4)2SO4	
0.25	g	Glucose	
10	ml	**Vitamin solution**	
20	ml	**Hutner's salts**	
0.071	g	Na2HPO4	
980	ml	Distilled water	
15	g/l	Agar, for solid medium	

1. pH 7
2. Make up the medium without the vitamin solution and autoclave it. The filter-sterilized vitamin solution is added to the cooled medium. The medium may be solidified by adding 15 g/l agar.


## Vitamin solution

0.1	mg	Vitamin B12	
2	mg	Biotin	
5	mg	Thiamine-HCl x 2 H2O	
5	mg	Calcium pantothenate	
2	mg	Folic acid	
5	mg	Riboflavin	
5	mg	Nicotinamide	
1000	ml	Distilled water	

1. Filter-sterilized


## Hutner's salts

10	g	Nitrilotriacetic acid (NTA)	
29.7	g	MgSO4 x 7 H2O	
3.335	g	CaCl2 x 2 H2O	
9.25	mg	(NH4)6Mo7O24 x 4 H2O	
99	mg	FeSO4 x 7 H2O	
50	ml	**"Metals 44"**	
950	ml	Distilled water	

1. Dissolve the nitrilotriacetic acid, adjust the pH to 7.0 with KOH (about 7.3 g). Dissolve other salts separately, combine and adjust the pH to 6.8 with NaOH or H2SO4.


## "Metals 44"

250	mg	Na-EDTA	
1095	mg	ZnSO4 x 7 H2O	
500	mg	FeSO4 x 7 H2O	
154	mg	MnSO4 x H2O	
39.2	mg	CuSO4 x 5 H2O	
24.8	mg	Co(NO3)2 x 6 H2O	
17.7	mg	Na2B4O7 x 10 H2O	
1000	ml	Distilled water	

1. Dissolve the EDTA and add a few drops of concentrated H2SO4 to retard precipitation of the heavy metal ions.